The overall goal of the proposed research is to adapt and apply an existing nucleic acid (NA) amplification, lab-on-a-card platform to identify enteric bacteria in patients presenting to primary health care settings or public health laboratories with symptoms of acute diarrhea. Component functions to be integrated on this platform include multiplex NA amplification and detection, sample processing to support direct use of clinical specimens, and dry reagent storage and handling. This new, robust, diagnostic test will provide a rapid, highly sensitive, specific, easy-to-use, cost-effective tool to distinguish bacteria likely used in bioterrorism attacks from bacteria that cause similar generalized symptoms. The four-year project is designed to achieve the following activities and milestones: 1. Develop immunological approaches to increase specificity and enable direct use of fresh stool or rectal swab specimens. 2. Develop a multiplex NA amplification, lab-on-card platform for rapid identification of Shigella dysenteriae, Shigella toxin-producing Escherichia coli, Campylobacter jejuni, and Salmonella species simultaneously from the same clinical specimen. 3. Optimize on-board reagent storage and handling capabilities. 4. Integrate reagent storage and handling, sample processing, and NA amplification and detection onto a single, lab-on-a-card, disposable enterics card system. 5. Collect clinical specimens to support research and development efforts and assessment of the clinical sensitivity and specificity of the assay. A consortium agreement among Program for Appropriate Technology in Health (PATH), Seattle, WA; Micronics Inc., Redmond, WA; Xtrana Inc. (formerly Biopool Inc.), Broomfield, CO; the University of Washington, Seattle, WA; and Washington University, St. Louis, MO, will provide for a world-class, multidisciplinary team skilled in microbiology; biochemistry; genomics; microfabrication; product development and scale-up of high-volume, regulated, disposable devices and instruments; DNA molecular physics; dry chemistry stabilization; clinical research; and overall technical management.